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Effect of Potassium Lactate and a Potassium Lactate-Sodium Diacetate Blend on Listeria Monocytogenes Growth in Modified Atmosphere Packaged Sliced Ham

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Effect of Potassium Lactate and a Potassium Lactate-Sodium Diacetate Blend on Listeria Monocytogenes Growth in Modified Atmosphere Packaged Sliced Ham 2297 Journal of Food Protection, Vol. 70, No. 10, 2007, Pages 2297-2305 Copyright ©, International Association for Food Protection Effect of Potassium Lactate and a Potassium Lactate-Sodium Diacetate Blend on Listeria monocytogenes Growth in Modified Atmosphere Packaged Sliced Ham L. A. MELLEFONT* AND T. ROSS Australian Food Safety Centre of Excellence, Tasmanian Institute of Agricultural Research, School of Agricultural Science, University of Tasmania, Private Bag 54, Hobart 7001, Tasmania, Australia MS 07-051: Received 30 January 2007/Accepted 9 May 2007 ABSTRACT Two commercially available organic acid salts, potassium lactate (PURASAL HiPure P) and a potassium lactate-sodium diacetate blend (PURASAL Opti.Form PD 4), were assessed as potential inhibitors of Listeria monocytogenes growth in modified atmosphere packaged (MAP) sliced ham in challenge studies. The influence of the initial inoculation level of L. monocytogenes (101 or 103 CPU g-I) and storage temperature (4 or g0C) was also examined. The addition of either organic acid salt to MAP sliced ham strongly inhibited the growth of L. monocytogenes during the normal shelf life of the product under ideal refrigeration conditions (4°C) and even under abusive temperature conditions (i.e., g0C). During the challenge studies and in the absence of either organic acid salt, L. monocytogenes numbers increased by l,OOO-fold after 20 days at goC and IO-fold after 42 days at 4°C. Both organic acid salt treatments were found to be listeriostatic rather than listericidal. The addition of either organic acid salt to the MAP ham also reduced the growth of indigenous microflora, i.e., aerobic microflora and lactic acid bacteria. The influence of these compounds on the risk of listeriosis in relation to product shelf life is discussed. Unlike most foodbome pathogens, Listeria monocyto­ modify or treat long shelf life refrigerated products in such genes can grow, albeit slowly, at refrigeration temperatures, a way that the growth of L. monocytogenes is prevented, at in the presence of >5% salt, and in the absence of oxygen least within the normal shelf life of the product. Organic (37). Processed meats, including ham and pate, are often acids and their salts, applied singly and in combination, packaged under vacuum or modified atmospheres and have been shown by many studies to prevent or greatly stored under refrigeration to extend their shelf life, typically retard the growth of L. monocytogenes in processed meats being 6 to 8 weeks or more. Although initial contamination under both recommended temperatures of storage (at or be­ with L. monocytogenes, when present, is usually low, i.e., low 4°C) and mild temperature abuse (e.g., up to 10°C). <10 CPU g-l (15, 28), its potential for growth in ready­ Generally, studies have focused on the activity of the so­ to-eat meats that are eaten without further cooking prior to dium salts (2-4, 9, 14, 17, 18,21,22,27,29,31, 34, 40), consumption is, prima facie, considerable. Moreover, it is but several also consider potassium lactate or potassium well documented that L. monocytogenes can grow in many lactate in combination with sodium diacetate (19, 26, 32, such products during refrigerated storage, and such foods 33). Most studies have considered the activity of organic have caused, or been strongly implicated in, outbreaks of acid salts in sausage products (e.g., bratwurst, frankfurters, foodbome listeriosis that have resulted in fatalities (6-8, 23, saveloys), but others have studied their effectiveness in 25). cooked ham (4, 34). Mbandi and Shelef (21) considered the L. monocytogenes present on raw ingredients can be effectiveness in sterile uncooked comminuted beef emul­ killed reliably by the heat treatments routinely applied dur­ sion. ing smallgoods processing. During slicing and packaging There are a range of commercial products available after processing, however, recontamination can occur (16, based on the salts of organic acids that are intended to pro­ 18, 24, 30, 39). During the past decade, the food industry vide protection against the growth of undesirable microor­ has implemented hygiene practices and technologies that ganisms on foods. This project was undertaken to assess have greatly reduced the prevalence of L. monocytogenes the antilisterial efficacy of two such products on an Aus­ tralian smallgoods product, specifically, PURASAL HiPure on ready-to-eat foods (20, 28). Nevertheless, the ecology P (potassium lactate) and PURASAL Opti.Form PD 4 (a of the organism, particularly its ability to colonize food potassium lactate-sodium diacetate blend). The influence, processing plants (38), still results in L. monocytogenes be­ and possible interaction, of the presence of either antilis­ ing detected, typically, in -1 to 5%, or more, of processed terial product, the initial level of L. monocytogenes, and the meat products (1, 5, 15, 20, 28, 41). temperature of storage on the development of populations One approach to minimize the risk of listeriosis is to of L. monocytogenes in modified atmosphere packaged * Author for correspondence. Tel: + 61 (0)3 62 266278. Fax: + 61 (0)3 (MAP) sliced ham are investigated. The influence of indig­ 62 267450; E-mail: lyndal.mel1efo~utas.edu.au. enous microbiota is also considered. 2298 MELLEFONT AND ROSS ~ J. Food Prot., Vol. 70, No. 10 MATERIALS AND METHODS aging. Uninoculated control samples were prepared by adding 0.1 ml of sterile pw. To minimize the potential for cross-contamina­ Ham preparation. Hams were prepared under commercial tion, these control samples were prepared prior to the samples conditions by a large Australian manufacturer. During processing, requiring inoculation with the L. monocytogenes. antilisterial treatments were applied to hams prior to cooking. The To preserve the integrity of the gas mixture, a self-adhesive treatments were as follows: the addition of 3% (wtJwt finished rubber septum was applied to each sample package, and the sy­ product) of PURASAL Opti.Form PD 4 (a blend consisting of ringe was inserted into the pack through the septum. An inoculum 54.5 to 57.5% potassium lactate and 3.7 to 4.3% sodium diacetate: volume of 0.1 ml was added to a comer of the base of the package Purac Asia Pacific Pte. Ltd., Singapore) or the addition of 3% (wtJ wt finished product) of PURASAL HiPure P (58 to 62% potas­ that did not contain any ham. The package was then inverted and sium lactate). Both products were provided to the smallgoods pro­ gently shaken (by hand) so that the slices of ham moved across ducer by Fibrisol Service Australia Pty. Ltd. (Heatherton, Victoria, the inner surfaces of the container and each other. This was done Australia). Finished hams were sliced by the manufacturer and to maximize the likelihood of even distribution of cells of L. packaged in -50-g lots into commercial thermoformed ridge bot­ monocytogenes or sterile diluent throughout the sample. Imme­ tom packs, composed of polyethylene teraphthalate and polyeth­ diately after inoculation (or addition of sterile PW), the septum ylene, and produced on horizontal form fill. The packages were was covered with self-adhesive tape. Inoculated samples were ap­ propriately labeled and incubated at either 4 or 8°C in walk-in then sealed under MAP (30% CO2 and 70% N2) . Control and treated samples were transported to the laboratory by commercial cool-rooms. The duration of storage was :5.76 and :5.57 days for transport operators under refrigeration, but no temperature records samples stored at 4 and 8°C, respectively. were available. Sampling. At appropriate time intervals, three samples from Bacterial strains. Five strains of L. monocytogenes were each treatment or control were assessed for levels of L. monocy­ used in combination for all "inoculated" samples. L. monocyto­ togenes, concentrations of lactic acid bacteria, and aerobic plate genes Scott A (type strain) and L5/22 (cold-smoked salmon iso­ count (APC). The entire sample from each package was asepti­ late) were obtained from the School of Agricultural Science, Uni­ cally removed and diluted I: I in PW and stomached for 2 min. versity of Tasmania. Strains 20425, 20432, and 20423, all envi­ Further W-fold dilutions in PW were prepared as required, and ronmental isolates from a smallgoods factory, were supplied by 100- or 250-111 aliquots were surface plated with a spiral plater Silliker Microtech Pty., Ltd., Melbourne, Victoria, Australia. (Autoplate 4000, Spiral Biotech Inc., Bethesda, Md.) onto agar Strains were maintained on bead suspensions in nutrient broth plates as follows. PALCAM plates (Oxoid CM877, and SRl50 (Oxoid CMl, Oxoid Ltd., Hampshire, UK) with 15% glycerol and antibiotic supplement) for L. monocytogenes were incubated aer­ stored at -70°C. obically at 37°C for 48 h. The deMan Rogosa Sharpe agar (Oxoid Preparation of inocula. Before each experiment, isolates CM361) plates for lactic acid bacteria were incubated aerobically were resuscitated from cryogenic storage by plating onto brain at 25°C for 72 h. Plate count agar plates (Oxoid CM463 APHA heart infusion agar (Oxoid CM225) and incubating at 25°C for 48 Standard Plate Count Agar) for APCs were incubated at 20°C for h. A primary culture of each strain was prepared by touching a 72 h. Colonies were counted manually, and log(viable cell count) sterile loop to five colonies and inoculating 10 ml of prewarmed of the triplicate samples was averaged (±standard deviation) and (25°C) tryptone soya broth (Oxoid CM129) containing 0.6% yeast plotted against time. The maximum sample volume plated was extract (TSBYE; Oxoid L21). The primary culture for each strain 250 111 of the I: 1 dilution on quadruplicate plates. This permitted was incubated without shaking at 37°C for 24 h. The primary a maximum test sensitivity of 2 CFU g-I. cultures were then serially diluted in 0.1% peptone water (PW; Water activity (Aqualab CX2, Decagon Devices, Pullman, Oxoid Bacteriological Peptone L37 containing 0.85% NaCl) and Wash.) and direct measurement of the pH (pH meter 250A.
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